Device and Method for Scanning Probe Microscopy

ABSTRACT

The invention relates to a device for scanning probe microscopy, said device comprising a scanning microscopy measuring device provided with a measuring probe for scanning microscopy measurements and a sample carrier for receiving a sample to be measured by scanning microscopy; a control device which is connected to the scanning microscopy measuring device in such a way that it is integrated into the system, and is designed in such a way as to automatically control the measuring device in order to perform a scanning microscopy measurement according to pre-defined control parameters; and/or an evaluation device that is connected to the scanning microscopy measuring device in such a way that it is integrated into the system, and is designed in such a way as to automatically evaluate measurements according to pre-defined evaluation parameters.

SCOPE OF THE INVENTION

The present invention encompasses, in general, apparatuses and methodsfor the investigation of biological systems, wherein such systems areinvolved which are enabled to be carried out by means of scanning probemicroscopic methods and by special applications of force-spectroscopicresearch, where scanning covers a raster patterned area.

BACKGROUND OF THE INVENTION

Biological systems and the processes which operate therein or are causedto so operate have their basic foundation on the interaction ofmolecules. Molecular forces in biological systems are different fromother molecular systems where chemical reactions and changes of physicalphases in a complete system are concerned. Statements about molecularinteractive activity in biological systems bring forward theadvantageous concept that such systems could be analyzed in order to beable in the future to formulate statements in greater detail.

For the measurement of molecular interchange in biological systems,among other methods, raster type scanning, based on probe centered,microscopic methods are in frequent use, in order to make determinationsregarding the topographic characteristics of surfaces by using highlateral and vertical resolution. In regard to “lateral resolution”, isto be understood that resolution in a single plane of a surface underinvestigation in a biological system is meant, while correspondingly,the resolution perpendicularly aligned thereto is designated as“vertical resolution”.

Examples for raster-scanning probe microscopic applications encompasssuch procedures as may be found in, for instance, SFM (scanning forcemicroscopy) or again in AFM (atomic-force microscopy).

With such scanning probe microscopy applications, it is possible, thatbesides the topology of a surface of a biological sample, also theelasticity thereof or the thereto applicable adhesion forces can bedetermined. The scanning probe microscopy, here normally referred to asa “force-spectroscopy”, determines molecular forces of the sample bymeans of an analytic probe which invades or reacts with the said sample,in order to quantitatively detect and characterize the interactiveexchange between individual molecules. Customarily, a probe includes ameasuring tip, which is carried on a freely extending arm or issupported in a cantilever fashion. When being used in an analysis, theprobe assembly is measurably caused to crisscross over the surface of asample, during which action, the lateral and vertical positions and/ordeflections of a probe can be recorded. Movements of a probe relative tothe sample are possible due to the elasticity of a probe itself andespecially the characteristics of the said cantilever holder thereof.Based on lateral and vertical positioning and/or deviations of a probe,molecular forces inherent in the sample and the therefrom inducedtopographical characteristics can be determined.

Customarily, movements of a probe are detected by optical measuringinstruments, wherein the resolution stands at about 0.1 nm and permit adetermination of forces in the range of a few pN (pico-Newton).

In order to determine the surface topography of a biological sample, thesurface of the sample and the probe of the said force-microscopicprocedure can be so mutually counter positioned, that a force, actingbetween them can be brought to a predetermined value (nominally about 50to 100 pN). Accordingly, a probe and the sample are so placed, relativeto one another, that a crisscross (raster) scan of the surface of thesample is enabled. In this way, the sample and/or the probe can movealso vertically, so that the forces acting between them are purposelyretained within the predetermined values. Movements of the sample andprobe relative to one another can be activated by means of apiezoelectric ceramic device.

One advantage of the scanning force-microscopy can be found therein, inthat biological samples in buffered solutions held at a relevantphysiological temperature (namely 4° C. and 60° C.) can be investigated.

FIG. 1 illustrates, in a simplified way, the principal of a scanningforce microscope. For the analysis of the surface of a biological sampleBP a probe S is employed. A probe S can be considered to be a probe-tipSp, which is suspended from a spring C. In a case of movement of a probeS and the biological sample BP relative to one another, (for examplealong the path W), dependent upon the respective surface topography, aprobe S and the sample SP are so mutually displaced in relation to oneanother in a vertically designated direction, that the displacement ofthe spring C remains constant.

At the present time, scanning probe microscopy, however, presents a taskwhich is time consuming and intensive of personal attention. Actually, agreat number of measurement increments, are necessary for a well foundedanalysis. For this reason, present applications of scanning probemicroscopy are entirely unsatisfactory, due to the small number ofmeasurements made, for instance, per day. Also the limited possibilitiesof the analyses, of the evaluation and the subsequent use of themeasured data, present considerable disadvantages.

PURPOSE OF THE INVENTION

The purpose of the invention is to make available a device and method,whereby the disadvantages of known scanned probe microscopyapplications, especially in regard to personal attention and timerequirements are set aside and the necessary measurements for a soundlybased investigation of biological samples become available. Further, thepresent invention is to enable that data acquired by raster scannedprobe microscopy methods can be put to use in an improved manner, whichexceeds in quality, that done within the state of the technology.

ABSTRACT OF THE INVENTION

For the achievement of the above stated purpose as well as for thecarrying out of the same, the present invention requires an apparatusfor raster scanned, probe microscopy as well as a method for theexecution of a scanning probe microscopy measurement in accord with theindependent claims 1 and 32. Advantageous embodiments become evident inthe subordinate claims, the description which follows and are shown inthe attached drawings.

The invented apparatus serves for scanning probe microscopy andencompasses a scanning microscopy measuring instrument. The saidmeasuring instrument includes a measuring probe for the said scanningprobe microscopy measurements and a probe carrier for the manipulationof a scanning probe in action. Further, a control device and/or anevaluation unit is provided.

The control device is integrated into the system, that is to say, is anintegrated component of the invented apparatus and bound to the rasteroriented scanning probe microscopic equipment. The control device isintended to regulate the said measuring device for the automaticexecution of a scanned probe microscopic analysis in accord withpredetermined control parameters.

Likewise, the evaluation unit is system integrated and bound to thescanning probe microscopy equipment. This said unit also forms anintegral part of the invented apparatus and is designed to automaticallyevaluate measurements acquired by means of the scanning probe microscopymethod in accord with the predetermined evaluation parameters.

With the invented apparatus, a physical setup is prepared for theautomatic carrying out of investigations of biological samples by meansof scanning probe microscopy methods and/or to execute and record theevaluations of the same. Further, the invented apparatus is to possessthe ability to cause the said measurements to be in accord with theinvestigative probe and to be expressed in terms compliant with thesystem employed, so that appropriate parameters for control andevaluation can be used. One advantage which arises from this arrangementis that such measurements can be acquired and analyzed without personalintervention throughout an entire day.

Advantageously, the control device and/or the evaluation unit isdesigned to recognize readings of the measurement equipment and/or theevaluation unit to obtain data for the determination of input parametersor parameter-sets of the measured values. This feature can also serve asa backup source for data available from measurements of the saidmeasurement and/or evaluation unit for control or evaluation parameters.

The identification of parameters or parameter-sets can be carried outunder the use of iterative “search algorithms”, i.e. algorithmscharacterized to identify specified expressions being searched for.Further, it is possible, that at least in regard to this feature, thecontrol device and the evaluation unit should be interconnected. In thecase of one embodiment example, the evaluation unit analyzes theforce-spectroscopic data during the course of an experiment. Upon theattainment of significant number of data inputs, the subsequentexperimental parameters were included and the experiment carriedforward. The experimental parameters can, in this way, be changed insuch a manner, that, for example, the changes in the measured forces,which changes were actually measured, (along with the attendant energiesand binding constants) can be determined with greater precision. In thisway, for example, it becomes possible for the researching person toautomatically examine interesting areas of molecular interaction.

This embodiment of the invented apparatus provides the possibility, aswill be more completely described in the following, that measurementsand samples can be classified, measurements on one probe can be comparedwith one another, optimized measurement strategies can be developedand/or immediately applied as well as the measurement strategiescurrently used can be optimized.

Advantageously, this apparatus encompasses a data storage memory for thestorage of the data acquired from the evaluation unit, which data arisefrom an evaluation of measurements by means of the rasterized scannedmicroscopic measurement device. This embodiment allows, for example, theconstruction of a data-bank with information including the measurementson samples which have been examined by rasterized scanning probemicroscopy. The advantage is, that with the said data storage memory, itis possible, during or following, a current series of measurements, datacan be retrieved for reference. The data storage memory can also be usedfor the retrieval of completed measurement results, whereby comparisonsmay be made with data currently being input to the said memory.

Advantageously, the data retrieval memory is designed in such a manner,that the respective conditions of measurement methods for the specifiedcontrol parameters and/or for the given evaluation parameters and/or forthe current measurement can be so captured, that an unambiguousinformation-linkage to the currently corresponding data from theevaluation unit can be achieved.

In the case of an advantageous embodiment, a probe possesses a resilientelement namely a spring or the like. The said resilient element canemploy an unrestricted, pivotal arm or a cantilever extension as itsactive structure. With this type of construction, further provision canbe made, that the forces at the measurement device, which act upon aprobe can be calculated.

In the case of a continued development of the invented embodiment, soprovided with the said resilient element, provision can now be made thatthe measurement device experiences alternating action between a probeand the sample on the basis of which, the forces acting upon a probe canbe determined by the application of an optical measurement system (forinstance, a laser beam deflection system, i.e. “beam bouncing”) and/orby means of piezo-electrical effects and/or the employment of magneticeffects arising from the said alternating motion of the raster action.

The invented apparatus can also possess a unit for the production of alight field and/or an electrical field and/or a magnetic field. It ispossible that the said fields can be static or dynamic fields, in whichinteractive cases a back-and-forth operation between the static and thedynamic effects can be proposed.

The said resilient element, which can advantageously be provided, couldbe a spring which would have a length within a range of 1 to 400micrometers, this being combined with or being itself an elastic orcantilever extension.

Further, provision has been made, that the control device would have thecapability of so regulating the resilient element, that the measuringprobe can be adjusted to a predetermined amplitude of vibration. Forexample, amplitudes in a range between 0.1 or 2000 nanometers could beselected.

Additionally, the invented apparatus can include a force producing unit,which can be assigned to the measurement device and/or to the controldevice.

In such an arrangement, the possibility exists that the control deviceis automatically able to so regulate the force producing unit, thatchanges for the active quality factors (Q-factors) of a probe can becarried out, so that corresponding forces can be directed to the saidresilient element.

An employment of the said force producing unit is especiallyadvantageously favored, if a probe is set into a predetermined vibratorystate. For the determination of the vibratory changes of the measuringprobe, it is possible to so adjust the evaluation unit to detect suchchanges in the form of resonance displacement and/or alterations inamplitude and/or phase shifting.

In the case of an additional embodiment, the measurement apparatuspossesses a probe positioning unit, which has the purpose of positioninga probe along all axes for translation or rotation of its occupiedspace. In this arrangement, provision has been made that the controldevice has the ability to automatically position and/or move the saidprobe by regulation of the said positioning unit for the attainment ofpredetermined probe positioning parameters.

In a case of the application of probe positioning unit, the proposal isto make use of probe positioning parameters, which include thefollowing:

-   -   movements of the measuring probe for the rasterized scanning of        the sample which has been placed on the sample-carrier, whereby        such movements embrace lateral displacements and/or        displacements in a range between 0.1 nanometers and a few        millimeters, preferably in a range between 0.1 nanometers and        500 micrometers,    -   movements of the measuring probe in a vertical direction,        whereby provision encompasses such displacements as might lie in        a range between 0.01 nanometers and 50 micrometers,    -   movements of the measuring probe in a vertical direction, where        concern is extended to a predetermined minimal separating        distance between the measuring probe and the sample, wherein the        possibility exists, that a separating distance regulation, for        example a PID and/or a phase-logic-regulator is used for the        control of the said movements,    -   a maximal duration of time for a contact of the measuring probe        with the sample which is situated on the sample-carrier,    -   a maximal repetitive frequency of contacts of the measuring        probe with the sample which is placed on the sample-carrier of        the said probe,    -   a maximal and/or an minimal measuring probe speed for the        movements of the measuring probe relative to the sample placed        on the sample-carrier of the said measuring probe,    -   a maximal and/or a minimal separating distance between the        measuring probe and the sample placed on the sample-carrier of        the said measuring probe,    -   a predetermined, force, which is held constant, and which acts        between the measuring probe and the sample placed on the        sample-carrier of the said probe, which, as an example, can lie        in a range between 0.1 and 3000 pN,    -   a maximal and or a minimal tension of the measuring probe, which        tension is exercised against the sample on the sample-carrier of        the said probe,    -   a maximal and/or a minimal compressive force of the measuring        probe, which force is exercised against the sample on the        sample-carrier of the said probe,    -   a maximal and/or a tensile force which acts upon the        sample-carrier of the said probe,    -   a maximal and/or a minimal compressive force change for the        compressive force of the measuring probe onto the sample-carrier        of the said probe,    -   a maximal or a minimal shearing force of the measuring probe        exercised against the sample which is on the sample-carrier,        and/or    -   a maximal or a minimal shearing force change rate for the shear        force exercised by the measuring probe against the sample placed        on the sample-carrier of the said measuring probe.

In the case of another embodiment a first detector unit is provided,which detector has the capability of determining the positions of aprobe, and/or the movements of a probe, favorably also the deviationsthereof, and/or, in a regular manner, also the forces acting on a probe,that is, regular in the manner of being repetitive with a frequency ofsome ten or some hundred kHz. In this arrangement, the control device,advantageously, is adjusted to automatically regulate the first detectorunit in accord with predetermined detection parameters.

Advantageously, the first detector unit encompasses position sensors todetermine position and or motion conditions of a probe. In the way ofexplanation, it is possible the LVDT-sensors, expansion measuringstrips, optical sensors, interference sensors, capacity sensors, can beused for this purpose. Particularly advantageous is an optical beamdiversion detector for the determination of the deflection of a probe.

Further, it is considered advantageous, that on the base of the firstdetector unit, immediately placed data on the positioning control and/orthe motion control of a probe and/or the sample under one or more closedregulation circuits is available. For this purpose, provision is made,that the first detector unit, at least in this consideration, isconnected with the control device.

Provision is also made, that the control device is enabled to controlthe first detector unit, in accord with predetermined detectorparameters, which include:

-   -   a preset detection rate, in respect to single, multiple and/or        all sizes to be captured, and/or    -   a sufficient limit of instances, within which the first detector        unit can carry out determinations of position, motion and/or        forces.

Advantageously, the evaluation unit has the ability to evaluate thevalues determined by the first detector unit. This can be done either byanalytical methods or in a statistical manner.

The evaluation unit can be so designed, that values obtained by thefirst detector unit can be classified. When this is carried out, it ispreferred, that classified values, that is the data available from theevaluation unit, which have been obtained as described above, bereturned into the measurement process, for example, in order to identifyspecial parameters sets of the measurement procedure.

In accord with an additional embodiment, the measurement apparatusincludes the sample-carrier positioning element, in order that thepositioning of the sample-carrier is precisely executed. In this way thecontrol device can be so adjusted, that the sample-carrier can beautomatically positioned by means of the regulation imposed by the saidsample-carrier positioning element. This regulation or control would becarried out in accord with preset sample-carrier positioning parameters,while the sample-carrier is being set in position or is being moved.

Advantageously, sample-carrier positioning parameters are to meet thefollowing requirements:

-   1) placement of the sample-carrier at the rastered point of the    sample affixed to the sample-carrier by means of a probe, whereby    lateral movements can be made and/or placement with a precision in a    range between 0.1 and 500 micrometers,-   2) a maximal time period for the duration of a contact of a sample    placed on the sample-carrier by the measuring probe,-   3) a maximum number of times for contacts of the sample placed on    the sample-carrier by a probe,-   4) a maximum and/or a minimum sample-carrier speed for movements of    the sample-carrier relative to the measuring probe,-   5) a maximum and/or a minimum separating distance between the sample    placed on the sample-carrier and a probe,-   6) a predetermined force, which is to be maintained as a constant,    which, for example is found in value to be between 0.1 and 3000 pN,    and which force acts between the sample placed on the sample-carrier    and the measuring probe,-   7) a maximum and/or a minimum tension exercised on the sample placed    on the sample-carrier by means of the measuring probe,-   8) a maximum and/or a minimum compressive force exercised on the    sample placed on the sample-carrier by means of the measuring probe,-   9) a maximum and/or a minimum tension force rate of change for a    tensile force acting upon the sample placed on the sample-carrier by    means of the measuring probe,-   10) a maximum and/or a minimum rate of change for a compressive    force acting upon the sample placed on the sample-carrier by means    of the measuring probe,-   11) a maximum or a minimum rate of change of a tensile force acting    upon the sample upon the sample-carrier by means of a probe, and    finally,-   12) a maximum and/or a minimum rate of change for a shearing force    acting upon the sample placed upon the sample-carrier by means of a    probe.

Advantageously, the sample-carrier positioning element can be a piezoelectrical actuator and/or a linearly active drive, which, for example,can be a voice-coil drive.

The above mentioned embodiments of the sample-carrier positioningelement are designed to produce a very precise positioning and/ormovements of the sample-carrier. The sample-carrier positioning elementcan also be so designed, that “rough” positioning and/or movements ofthe sample-carrier become possible. This would be true particularly in arange between 100 nm and 30 cm. The advantage of such the sample-carrierpositioning element, having, as said, “rough” positioning, is to befound in that previously determined positioning and larger movements canbe quickly carried out. Exact positioning and movements can then besubsequently applied.

Advantageously, positional and/or movement detecting sensors can beincluded in the said sample-carrier positioning elements, which sensors,for example, by means of a closed regulation circuit can make availableinformation for control of the sample-carrier positioning element.During such an operation, the above mentioned position sensors and/oradditional sensors can be employed.

Advantageously, the measuring system includes the sample chamber, intowhich a liquid is introduced, with which liquid the sample to be placedon the sample-carrier is to be immersed. In this matter, where“immersed” is concerned, one is to understand, that at least the area ofthe sample, which is under investigation, is immersed in fluid, forinstance, at least a certain surface of the sample is to be so wetted.In this way, the control device is enabled to monitor or to be adjustedfor, the individual characteristics of the given parameters of the fluidso employed.

As fluid parameters can be named:

-   -   a predetermined temperature,    -   a predetermined temperature curve,    -   a predetermined pH value,    -   a predetermined pH value curve    -   a predetermined electrolyte content,    -   a predetermined electrolyte content curve,    -   a predetermined flow,    -   a predetermined change of flow,    -   a predetermined volume level of flow, and/or    -   a predetermined number of biological and/or chemical markers or        properties.

By means of the use of a predetermined number of biological or chemicalproperties as fluid parameters, it is possible that a number thereof canbe selected for use as fluorescence markers and or radioactive markers.Advantageously, markers are employed which function in chemical orbiological manners and can exhibit selective characteristics of thesample to be investigated.

In the case of an additional embodiment, feeding equipment is provided,by means of which a fluid, which is to be added (for example, a fluid orfluids such as a buffer or a reagent) can be made available. With thisfeature, the control device can be designed in such a way, that the saidfeeding equipment is automatically supervised and, if required, can beso controlled, that for the selected auxiliary fluid predetermined sideeffects, preferably even their desired compositions, can be broughtabout as a continuing condition in the sample chamber. In the way ofexample, reagents can be automatically combined with one another, inorder that fluid parameters such as the pH value and the like can beattained in the final mix.

Advantageously, the feeding equipment allows the entry of fluid to thesample-carrier in that area, in which during the measurement procedure,the sample is placed. Further, again in a favorable manner, thearrangement allows that the feeding equipment is also automaticallycontrolled by the control unit.

The feeding equipment can include as its active agent, for instance, anindividual or a multichannel pump.

For the monitoring of the fluid in the sample chamber and/or to aid inthe current side effects of a given fluid, a second detector unit can besupplied. In this case, to react to the level of fluid registered by thesecond detector unit, the first feeding equipment can be automaticallycontrolled.

Advantageously, a temperature enclosure is included in the analyticsystem. This embraces at least both a probe and the sample-carrier. Inaddition, provision is made so that the temperature enclosure is also anadditional component of the measurement equipment, such as, for example,if one be available, the force producing means, a probe positioningunit, the sample-carrying unit positioning element, the sample chamber,the first feeding equipment and/or the second feeding equipment, whichis described in the following. In the case of the second feedingequipment, advantageously the control of the temperature enclosure bymeans of the control device is carried out in accord with the previouslydescribed methods shown above.

Advantageously, in this arrangement, provision is further made, that thecontrol device is so enabled to regulate the temperature enclosure, thata predetermined temperature is maintained therein and/or at least apredetermined curve of temperature change will be followed. For example,it is possible, that the control of the temperature enclosure is socarried out, that the duration of constantly maintained temperature canbe replaced by other durations, in order that it may coincide withdesired temperature curves.

Especially, preference is given to the fact that the raster microscopicmeasurement equipment is a force microscopic measurement equipment,advantageously for the recording of force separation curves. From this,it is possible that information regarding interactive activity andconnection forces of individual molecules can be achieved.

Further advantage is gained, in that the measurement apparatusencompasses, in addition to the above, an optical detector unit,preferably such a device based on fluorescence and/or transmitted lightconducting microscopic technology (such as DIC and/or phase contrast, ofa bright or dark field technique).

The above purpose can also be achieved by means of the inventedprocedure for the execution of a raster probe microscopic measurement.The invented procedure, as well as the preferred embodiment examples ofthe same are defined in the claims designated for the said procedure.

BRIEF DESCRIPTION OF THE DRAWINGS

In the following description, preferred embodiment examples are used asexplanatory models for the attached drawings. There is shown in:

FIG. 1 a schematic presentation of the principle of a raster forcemicroscope (AFM)

FIG. 2 a schematic illustration of an embodiment example 1 of theinvented apparatus for raster probe microscopy, and

FIG. 3 a representation of a probe placement intended for use in theinvented apparatus.

DESCRIPTION OF ADVANTAGEOUS EMBODIMENT EXAMPLES

As is shown in FIG. 2, the embodiment example 1 embraces a raster probemicroscopic measuring apparatus 2, exhibited, somewhat, as anatomic-force, microscopic measurement apparatus (AFM) which includes inits composition a measuring probe 4 for raster force microscopic orforce microscopic measurements and the sample-carrier 6, upon whichsample-carrier a biological sample 8 may be placed for the carrying outof an analysis.

The measuring probe 4 is designed as a cantilever extension, whichcantilever serves to allow resilient properties to the said probe 4. Atthe free end thereof, in particular, at the tip of the cantilever aprobe (not shown) can be integrated as a unified component. Thepresentation in FIG. 2 principally shows a probe. In any case, thisprobe is shown as “probe 4” to be used within the measurement apparatus2.

In particular, the measurement apparatus 2 can possess two, four, six oreight probes. The number of probes 4 can extend itself even to 100 ormore. To incorporate this extended number of probes 4 to be used withinthe measurement apparatus recourse may be made to the use of theso-called “cantilever chips” (see FIG. 3).

FIG. 3 shows an electron-microscopic photograph of a set of cantileverchips, which pictures eight spring-like units, 4 ₁ to 4 ₈ in that typeof construction. Within the framework of nanotechnological installationpractices, such chips supported by a hundred or more cantilevers arepossible.

Giving consideration to the sample to be analyzed and relative to thecurrent analysis being carried out, it can be necessary to set a probeinto vibration at a predetermined amplitude. For this reason, themeasurement apparatus includes a force producing unit which coacts withthe measuring probe 4 and with the resilient unit. This force producingunit is not illustrated. The force producing unit, however enables themeasuring probe to move in such a manner, that a desired quality-factor(hereinafter “Q-factor”) appropriate to the analysis can be achieved.

Additionally, with measurement apparatus possesses the samplepositioning device 18 (not shown in detail), which serves the purpose ofpositioning a probe 4 in relation to the sample 8. Relative to thecharacteristics of the sample 8 to be analyzed, as well as relative tothe analysis itself, it is possible that by means of the samplepositioning device 18, a probe 4 can assume a correspondinglyappropriate position relative to the sample 8 and likewise follow adesired path thereon.

By means of a first detector unit 12, which is included in the assemblyof the measurement apparatus 2 or, otherwise, is assigned thereto, it ispossible that the positions and travel paths of the sample 8 can bedetermined. The first detector unit 12 is made as an optically baseddetector and encompasses a source of radiation 14, which can be a laserbeam emitter, and includes further a light first receiver 16 whichcollects the light from the said radiation source 14 after itsinteraction action with a probe 4. The said radiation source 14 targetsa probe 4, particularly in the area inclusive of the free ends thereof,while interactive actions, specifically here reflections of the lightfrom the light source 14, are collected by a second receiver 16. Thesecollected reflected light rays are then conducted to the evaluation unit42 for the determination of positioning and movements of the measuringprobe 4.

The measuring apparatus 2 possesses further the sample-carrierpositioning element 10, which allows the said sample-carrier positioningelement to be integrated in, and be come available to, the saidmeasuring apparatus 2, thus being a component thereof The sample-carrier6 and the sample-carrier positioning unit 18 can also be designed a asseparate and discrete components.

The sample-carrier positioning unit 18 here provided is actuated by apiezo-electrical electrical element, which energizes the movements andpositioning of the sample-carrier 6 and therewith the thereonsuperimposed measuring probe 8 in its relative space.

The sample chamber 20 encapsulates the measuring probe 4 and the sample8 or encloses at least that area of the sample 8 which is to beanalyzed, for example, this said area can be the upper surface thereof.The sample chamber 20 is closed in a fluid tight manner, absent the inand out lines, which lines are described in the following, and on thisaccount presents a closed-off space in relation to its ambientsurroundings. The sample chamber 20 can be totally dedicated to theacceptance of incoming fluid or as is shown in FIG. 2, a fluid chamber22 can be provided, which is so designed, that the measuring probe 4 andthe sample 8 or more precisely, those areas thereof which are to beanalyzed, are immersed in fluid.

By means of a second detector unit 24, the level of fluid in the samplechamber 20 can be determined. The second detector unit 24 is designed asan optical measuring instrument, which includes in its assembly a lightsource 26, for instance, this being a laser beam emitter and a receiver28. The light from the said laser source 26, after its interaction withthe fluid in the sample chamber 20 is collected by the receiver 28.Readings in regard to the level of the fluid in the sample chamber 20,which readings are activated by the received laser light are possible toobtain.

The sample chamber 20 is connected with a feed line 30 by means ofwhich, fluid can be introduced into the sample chamber 20. By means of adischarge line 32, fluid from the sample chamber 20 finds an outlet fromthe said sample chamber 20.

The input side of feed line 30 is connected with a feed unit 34, bymeans of which the individual, combined, mixed and blended fluids of theinput line 30 can be introduced into the sample chamber 20. The feedunit 34, for this purpose, includes one or more pumps or multichamberpumps and apparatuses (not shown) for the mixing and blending of thesaid fluid mixtures. By means of the fluid supply lines 36 ₁ . . . 36_(n) the feed unit 34 obtains various fluids from their appropriatereservoirs (not shown). In the case of embodiment 1, which is the totaldiagram of FIG. 2, by means of the supply unit 34 fluids, which aredesired, and so approved by an analysis of the sample 8 and/or necessaryfluids are introduced, such as, for example buffer solutions and reagentmaterials.

The measurement apparatus 2 and the additional, above describedcomponents of the embodiment—absent the fluid sources connected with thesupply lines 36 ₁ . . . 36 _(n) and parts of the supply lines 36 ₁ . . .36 _(n)—are housed in a temperature controlling enclosure 38. Thistemperature enclosure 38 possesses, at least where temperature isconcerned, a temperature directed closure against the ambientenvironmental conditions.

The embodiment form 40 serves for the control of the components whichfind themselves housed within the temperature enclosure as well as theconditions of the temperature enclosure itself. In FIG. 2 are presentedthe line connections 44 and 46 which represent required interconnectionsbetween the control device 40 and the feed unit 34, as well asconnections for sample-carrier positioning element 18.

The evaluation unit 42 contains principally all components which areconfined within the temperature chamber 38, insofar as such componentsare designed to be so enclosed. Further, the said evaluation unit alsoobtains from the temperature enclosure 38, data, measurement signals,and the like, in order to transmit and evaluate the actual operationalconditions, i.e., the actual results, obtained from the saidmeasurements. Representing the connections which permit the aboveoperation, FIG. 2 shows connection lines 48, 50 and 52 between thedevaluation unit 42 and the feed unit 34, the sample-carrier positioningelement 18 and the second detector unit 24.

Further, the embodiment designated as 1 encompasses a data storagememory 54, which is connected to the control device 40 and theevaluation unit 42. The data storage memory 54 serves for the storage ofdata, measurement signals, and the like, which have been obtained fromthe evaluation unit. These data are available from the evaluation unit42 along with parameters, which can be used by the control device 40 forthe regulation of the said embodiment 1. These and other data are sostored, in accord with the following described information:

In particular, the data storage apparatus 54 is so designed, that it mayalso be employed as a data bank, in which storage is provided foralready evaluated data and for outside data which has a relation to theembodiment 1.

In the case of the execution of a raster probe microscopic measurement,the control unit regulates, besides the true force-spectroscopicexperiment, also all experimental conditions, such as temperature, theactive pH value of the sample 8, the coacting electrolyte(s) of thesample 8 as well as the supply of pharmaceutical, biochemical andchemical additives. Further the control device monitors the controldevice 40 for predetermined operational points of time, also checks theagain predetermined duration periods or active parameters for theon-going measurement and also supervises the side conditions and finallycontrols the measurement in such a manner, that specifications intendedfor that measurement are held.

The evaluation unit 42 analyzes the force-spectrum acquired by a probe 4during the investigation in regard to sample 8 and is able to interpretthis force-spectrum. Relative to this operation, the possibility alsoexists, that for example, upon the attainment of a preset value, themeasurement then in course can be brought to a termination and a newmeasurement initiated with new specifications (for instance, ambientcondition changes). The control device 40 and the available data fromthe evaluation unit 42 also enable iterative measurement cycles beingcarried out, in order that concurrent reactive effects are determined,which could enhance specified interactive operations.

During the measurement, a probe 4 is in a crisscrossing relationshipwith the sample 8, whereby, in the presence of interactions between aprobe 4 and the sample 8, (biological) molecules situated on the surfaceof the sample 8 can be detected. Relative to the formulation of themeasuring probe 4 and the type of sample 8, defined contact durationtimes and/or frequencies of contacts between a probe 4 and the sampleare required. These parameters are applied and adjusted by the controldevice 40, which also supervises and if necessary corrects the ongoingprocedure. In this matter, it can be advantageous, especially in theexecution of a completely automatic measurement, to prepare the sample 8to an optimum condition. More detailed methods for this preparation areto be found in the following description.

During the measurement procedure, the control device 40 regulates aplurality of relevant experimental conditions, such as, for example,maximum/minimum tension and compressive forces between a probe 4 and thesample 8, also the speeds with which the relative movements between aprobe 4 and the sample 8 are carried out, likewise, the number ofmeasurement points (resolution) and the maximum/minimum separatingdistances between a probe 4 and the sample 8.

As this is done, it is possible to proceed with the measurements, bymeans of which individual, several or all experimental conditions areheld constant and/or may be systematically and/or chaotically changed.Thus it is, for example, possible to execute a measurement in which,except for the travel speed of a probe 4, all experimental conditionsare held constant.

As soon as a predetermined number (for example, a thousand) ofmeasurements have been undergone by a single sample, it is possible tosend the acquired data to a data set. Additional data sets can be madeby means of changed experimental conditions, and then comparisons can becarried out. This makes it possible to analyze different biologicaland/or medicinal relevant experimental conditions in regard to theirinfluence on molecular interactivity.

A given value, which can influence the analyses of biological samples,would be thermal alterations.

The said temperature enclosure 38 is provided in order that thermaldrift during experimental conditions can be minimized. This might be,for example, changes arising from buffers coacting with sample 8.Further it is possible that a heating or a cooling element (forinstance, a Peltier-element) can be employed, in order that thetemperature of the sample 8 itself can be controlled. Such aheating/cooling element can, for example, be placed under thesample-carrier 6.

Bimolecular interactivities are, as a rule, very dependent upon thegoverning, physiological ambient conditions of the measurement. These,on this account, should be monitored during a measurement procedure andaccordingly provided with control. In this way, during the measurement,the desired surrounding conditions are maintained, which, generally,stimulates the inherent properties of the sample 8. Thus, for example,provision is made, that during a measurement procedure, the liquid levelof a buffer solution in the sample chamber 20 is monitored andcontrolled by means of the second detection unit 24 at predeterminedintervals or is supervised continually and, if necessary, the supplyequipment 34 is so operated, that a desired liquid level is correctlyheld or can be properly reached. In this way, it is possible, thatduring a measurement procedure a measurement of a lessening of thebuffer level due to evaporation can be compensated for. In this wayalso, pH-variations as well as changes regarding the employedelectrolyte or other materials which can interact with the sample 8 canbe placed under monitoring supervision and, if required, alsocontrolled. By means of an intended control of buffer solutions in thesample chamber 20, evaporation losses and salting-out occurrences can beavoided.

In the control of buffer solutions which are in the sample chamber 20,the possibility exists that fluid movements, that is, for example, aturbulence or swirling may occur, which can influence the accuracy ofthe measurement. For instance, fluid movements in the sample chamber 20can activate vibratory resonances in the measuring probe 4. In order toprevent this, provision is made, to the effect that the control device40 interrupts a procedure which is already in operation, if suchdisturbances are detected and/or predicted.

The speed, with which a measurement can be carried out with, plays anessential role. In any case, quickly executed measurement procedures aresubject to question. In order to achieve a high degree of quality and atthe same time perform a measurement in a short time, the inventionallows, that the measuring probe 4 can be moved with a greater thannormal speed and likewise quickly assume a desired position. When thisoccurs, then provision is made, that during a measurement procedure, asmeasurements are taken at different and/or the same points, differentspeeds for the movement and/or positioning of a probe 4 may be employed.This control of varying speeds and/or positioning enables, in the caseof force spectra (i.e. curves of force-intervals) permits theacquisition of detailed, automatic reporting in regard to molecularinteractions. In addition in this way, measurements are optimized, inthat the resolution, with which the force-spectra were acquired, isincreased. This situation can be achieved, in that even the smallestforce, which lies within the detection capability of a probe 4, can beminimized. The smallest detectable force depends on, among otherconditions, the resilient properties of the measuring probe 4. In orderto capture the smallest possible, detectable force, it is advisable tomake use of such probes, which exhibit the greatest degree ofspring-related characteristics and which show a highresonance-frequency.

A further possibility exists, in increasing the tractive velocity of aprobe 4. High speeds of movement can induce special hydrodynamicturbulences, which in turn create undesirable displacements of themeasuring probe 4. As a result, in a case of selected high speedoperations, undesirable noise reactions can be infused into measurementdata, whereby the sensitivity for the said lesser forces between a probeand the sample 8 is diminished. This disadvantage can be avoided, by theuse of probes, which possess the shortest possible length along with aminimum expanse of elasticity therein. Such probes exhibit, incomparison with conventional probes, a clearly superior hydrodynamiccharacter and permit obviously increased tractive speeds. In any case,such probes deflect from their chosen paths to a lesser extent that theconventional probe. On this account, the detector unit 12, in theinvented case, can be designed as the greatest possible, opticalassembly with special optical features.

In order to carry out raster probe, microscopic measurement along withstatic measurements, a probe 4, during the operation of a measurement,is set into vibrations of low amplitude, namely 0.1 to 10 nm. Adifficulty related to dynamic raster probe, microscopic measures, can befound therein, in that governing low quality factors (Q-factors), whichcan be attributed to measuring probes 4 which are immersed in puffercontaining solutions. Normally, molecular interactions can be capturedby means of resonance displacement, possibly at the maximum possiblevalue thereof and these can be determined by a probe. The resonancecharacteristics of a probe are proportional to the Q-factor, whereby alesser Q-factor can lead to a broadly spread resonance maximum. On thisaccount, a reduced sensitivity of the detection of force undercircumstances of a reduced Q-factor can be brought about. Additionally,such characteristics evoked by (for example) hydrodynamic turbulence offluid surrounding a probe or by the elastic action of a probe itself,(for instance, damping means) can contribute to dissipative interactionsin the analysis. Accordingly, in the case of embodiment 1 (see FIG. 2),provision has been made to increase to increase the Q-factor in such away, that by means of a positive feedback loop, an external force isapplied to a probe 4. Thereby, it is possible, that the Q-factor can beimproved by at least three times and often many time more, whereby thesensibility to force lies in the range of a few pN.

To obtain an automatic analysis from the results of measurement,provision is made, that during a measurement procedure, individualforce-curves are made under the use of the evaluation unit 42. As far asobtaining statistical analyses, the procedure would be as follows:

To begin with, the force-curves are so compensated among themselves,that they can be compared with one another. This can be done, forexample, by establishing a common unit separation along the zero line(reference value or abscissa) and/or by accordingly extending orcompressing the given curves to match.

Thereafter, it is possible the entire measurement procedure can bestatistically analyzed, in order that a determinative view may beobtained regarding proteins into the forces necessary for the probableapportioned degree of individual processes for folding and/or unfolding.

In addition, it is possible that the force-curves can be classified,viewed in real or supposed superimposition, and so determined as tocharacteristics. For the classification of force-curves, for example thelength of the individual force-curve can be seen, and number andposition of the therewith evident maximum forces can be determined. Thelength of a force-curve discloses the directions in which the expectedinteractions proceed. The number and positioning of force-maxima permitstatements to be made in regard to the collective and/or individualresults of interaction procedures. By means of a classification of forcecurves, it is possible that data, especially graphically illustrateddata, can be obtained in regard to different interaction processes.

By means of an overview of class of classified force-spectra, the noiseof the individual curves is reduced. Thereby the actual interactiveprocedures, which are illustratively disclosed by the force-curves,become obvious to the observer. In addition, statements about possiblevariations of the interactive procedures can be made, which statementsare based on standard deviations from already determined force-curves.

Comparisons of interactive procedures, carried out at the sameexperimental conditions on samples under the same experimentalconditions are made possible, when the following conditions are present:

-   a) a classification of force-curves,-   b) a determination of force-curves of a common class,-   c) interaction procedures which are related to or similar to one    another, and-   d) and a subsequent statistic analysis.

In this way, concerning the embodiment 1, three mutants of the samereceptor, which differentiate themselves from one another inpoint-mutations, are identified by means of their interactive spectraand can be compared with one another. Also statements in regard to theeffects of an input of mutations into local interactions of a protein aswell as the interaction of the given protein or proteins with othermolecules can be carried out.

By means of the data storage memory 54, it becomes possible to create adata-bank for force-spectra, in order, for example, to characterizetypical interactive procedures of various samples under differentexperimental conditions. For access to a data bank of the data storagememory 54, provision is made to employ different search strategies. Forexample, the structure data of an unfolded protein can be used, in orderthat structurally related proteins can be localized and thecharacteristics of their unfolded outline can be compared. In order tocompare different unfolded spectra, it is possible, by means of theevaluation unit 42, to evaluate data in the said data memory bank, inorder that, for example, different force-spectra to superimpose on oneanother and thus to compare. This makes it possible, to make statementsin regard to dependencies of interaction procedures where experimentalconditions are concerned. Furthermore, interactive procedures ofdifferent samples can be judged, as to whether or not the stored dataindicates that their interactive procedures are comparative, similar orhave the same characteristics.

Further, provision has been made, that that databank access can be madein relation to the presence of forces, physiological dependencies,interactive spectra and experimental conditions.

1. A device for raster probe microscopy with: a raster microscopicmeasuring apparatus, which possesses a measuring probe for rastermicroscopic measurements and a sample-carrier for the positioning of asample, which sample is to be subjected to raster microscopic analysis,a control unit which is systematically integrated into the rastermicroscopic measuring apparatus, whereby the said control unit isdesigned to regulate the measuring apparatus in the execution of anautomatic raster microscopic procedure in accord with predeterminedcontrol parameters, and an evaluation unit which is systematicallyintegrated with the microscopic measuring apparatus, whereby theevaluation unit is designed to evaluate measurements from the automaticraster microscopic procedure in accord with predetermined evaluationparameters. 2.-31. (canceled)
 32. A procedure for the execution of araster probe microscopic measurement consisting of the following steps:the predetermination of control parameters and/or evaluation parametersfor measurement by the raster microscopic method and the placement of asample to be measured by the raster microscopic method onto asample-carrier positioned on a measurement apparatus for the use of aprobe of the said measurement apparatus, whereby, the measurementapparatus, for the carrying out of the raster microscopic measurement isautomatically regulated in accord with the predetermined controlparameters by a control unit, and/or the raster microscopic measurementis automatically evaluated in accord with the predetermined evaluationparameters by an evaluation unit. 33.-54. (canceled)
 55. An apparatus inaccord with claim 1, wherein the control unit and/or the evaluation unitare so designed for the purpose of receiving and retaining data andemploying the said data for the determination of control and/orevaluation parameters and producing a continuous display thereof.
 56. Anapparatus in accord with claim 1, wherein a data storage memory isprovided for a retention of data produced from the evaluation unit byits evaluation of measurements made by means of the raster microscopicmeasurement apparatus.
 57. An apparatus in accord with claim 56, whereinthe data storage memory for a retention of predetermined controlparameters and/or data acquired by the measurement under conditions ofgiven measurement is designed in accordance with receiving correspondingdata made available by the evaluation unit.
 58. An apparatus in accordwith claim 1, wherein the probe comprises a resilient unit, and whereinthe evaluation unit is designed to evaluate the forces which act uponthe probe.
 59. An apparatus in accord with claim 58, wherein the probeis designed to accept signals of interaction between the probe and asample by means of an optical measurement system and/or by means ofpiezoelectric effects and/or by means of magnetic interactive responses.60. An apparatus in accord with claim 58, which possesses a unit for theproduction of one or more fields to which the probe can react, suchfields namely, light, electric or magnetic fields.
 61. An apparatus inaccord with claim 60, wherein the field producing unit is enabled toproduce statistic and/or dynamic fields.
 62. An apparatus in accord withclaim 58, wherein the resilient unit comprises a spring and/or acantilever extension.
 63. An apparatus in accord with claim 58, whereinthe control unit is designed to control the measuring apparatus in sucha manner that the probe is subjected to vibration having a predeterminedamplitude.
 64. An apparatus in accord with claim 58, wherein theapparatus possesses a force producing element which coacts with theresilient unit, and the control unit automatically controls the saidforce producing element to regulate the Q-factor of the probe.
 65. Anapparatus in accord with claim 63 wherein the control unit recognizeschanges of oscillation in the form of resonance shifts and/or amplitudechanges of the measuring probe.
 66. An apparatus in accord with claim 1,wherein the measuring apparatus includes a probe-positioning unit forthe placement of the probe in all translation and rotation effects ofthe operative space, and wherein the control unit controls theprobe-positioning unit to automatically position or move the probe inkeeping with predetermined probe positioning parameters.
 67. Anapparatus in accord with claim 66, wherein the control unit is sodesigned, in that the probe positioning unit is to carry out itsfunction within the bounds of predetermined probe positioning parametersof the control parameter group, which embrace: movements of the probefor raster apportioned scanning of a sample placed upon thesample-carrier, movement of the probe in a vertical direction, movementof the probe in a vertical direction relative to a predetermined minimalseparating distance between the probe and a sample, a maximum period oftime for a contact of the probe with a sample placed upon thesample-carrier, a maximum number of contacts of the probe with a sampleplaced upon the sample-carrier, a maximum and/or a minimum probe speedin the movements of the probe relative to a sample placed upon thesample-carrier, a constant probe speed, a constant change of probespeed, a maximum and/or a minimum separating distance between the probeand a sample placed upon the sample-carrier, a predetermined, constantlymaintained, retention-force between the probe and a sample placed on thesample-carrier, a maximum and/or a minimum tensile force of the probeacting upon a sample placed upon the sample-carrier, a maximum and/or aminimum compressive force of the probe acting upon a sample placed uponthe sample-carrier, a maximum and/or a minimum rate of change of tensionforce applied by the probe to a sample placed on the sample-carrier, amaximum and/or a minimum rate of change of compression force applied bythe probe to a sample placed upon the sample-carrier, a maximum and/or aminimum shearing force applied by the probe upon a sample placed uponthe sample-carrier, and/or a maximum and/or a minimal shearing force orrate of shearing force applied by the probe upon a sample placed uponthe sample-carrier.
 68. An apparatus in accord with claim 1 with: afirst detector unit for the detection of positions of the probe and/ormovements of the probe and/or forces effecting the said probe, wherebythe control apparatus is designed to automatically regulate the firstdetector unit in accord with predetermined detection parameters.
 69. Anapparatus in accord with claim 68, wherein the first detector unitincludes position sensors for the capture of positions and/or movementsof the probe.
 70. An apparatus in accord with claim 68 wherein: thecontrol unit is designed to regulate the first detection unit in accordwith predetermined detection parameters of the control parameter group,which encompass: a predetermined rate of detection, an occurrencefrequency, with which the detection actions for positions of the probeand/or motions of the probe and/or effective forces being applied to theprobe are carried out.
 71. An apparatus in accord with claim 68 whereinthe evaluation unit is designed to automatically evaluate positionsand/or movements and/or forces captured by the first detector unit. 72.An apparatus in accord with claim 68 wherein the evaluation unit isdesigned to classify the positions and/or movements and/or forcescaptured by the first detector unit.
 73. An apparatus in accord withclaim 1 wherein the evaluation unit comprises a sample-carrierpositioning unit for the positioning of the sample-carrier, and whereasthe control unit is designed to automatically position and/or move thesaid sample-carrier by means of regulation of the sample-carrierpositioning unit in accord with predetermined sample-carrier positioningunit parameters.
 74. An apparatus in accord with claim 73, wherein thecontrol unit is designed to regulate the sample-carrier positioning unitin terms of predetermined sample-carrier positioning unit parameters ofthe predetermined control parameter group which encompasses thefollowing: movements of the sample-carrier at rastered scanning of thesample placed upon the said sample-carrier, which scanning is performedby means of the probe, a maximum length of time for a contact between asample placed on the sample-carrier and the probe, a maximum number ofcontacts between a sample placed on the sample-carrier and the probe, amaximum and/or a minimum sample-carrier speed for movements of thesample-carrier relative to the probe, a maximum and/or a minimumseparating distance between a sample placed on the sample-carrier andthe probe, a force, which is to be maintained at a constant value,acting between a sample placed on the sample-carrier and the probe, amaximum or a minimum tensional force exercised on a sample placed on thesample-carrier, which force emanates from the probe, a maximum and/or aminimum compressive force exercised on a sample placed on thesample-carrier, which force emanates from the probe, a maximum and/or aminimum rate of change of the tensile force for the tensile force actingon a sample placed on the sample-carrier, which tensile force isexercised by the probe, a maximum and/or a minimum rate of change of acompressive force for the compressive force acting upon a sample whichhas been placed on the sample-carrier, which compressive force emanatesfrom the probe, a maximum and/or a minimum shear-force acting upon asample placed on the sample-carrier caused by a shearing-force exercisedby the probe, and/or a maximum and/or a minimum rate of change of ashear force acting upon a sample placed upon the sample-carrier, whereinsaid shearing force emanates from the probe.
 75. An apparatus in accordwith claim 73, wherein the sample-carrier positioning unit possesses apiezo electric actuator and/or a linear drive path.
 76. An apparatus inaccord with claim 1 wherein: the measuring apparatus comprises a samplechamber for the acceptance of provided fluids into which a sample placedon the sample-carrier is to be immersed, and the control unit isdesigned to control predetermined fluid parameters for the fluid.
 77. Anapparatus in accord with claim 76, wherein the control unit is designedto regulate, within the terms of predetermined fluid parameters of thecontrol parameter group, which encompass: a predetermined temperature, apredetermined temperature curve, a predetermined pH value, apredetermined pH curve, a predetermined electrolyte, a predeterminedelectrolyte curve, a predetermined flow, a predetermined flow curve, apredetermined level of fluid, and a predetermined quantity of biologicaland/or chemical identifying features.
 78. An apparatus in accord withclaim 1 which comprises a feeding unit for the flow of fluid to a samplechamber, wherein: the control unit is designed to control the saidfeeding unit in such a manner, that predetermined side effects of thefluid in the sample chamber are retained.
 79. An apparatus in accordwith claim 78, wherein the feeding unit comprises a pump and/or amultichamber pump.
 80. An apparatus in accord with claim 78 wherein thefeeding unit comprises a fluid level device for the detection of fluidlevel in the sample chamber, and the control unit is designed toautomatically regulate the feeding unit, in response to a signal fromthe fluid level device regarding fluid level.
 81. An apparatus in accordwith claim 1 comprising a temperature enclosure which encases the probeand the sample-carrier, and the control unit is designed to govern thetemperature enclosure in keeping with predetermined temperatureparameters.
 82. An apparatus in accord with claim 81 wherein the controlunit is designed to so govern the temperature enclosure, that apredetermined temperature is maintained and in that a predeterminedtemperature curve is followed.
 83. An apparatus in accord with claim 1wherein the raster microscopic measurement apparatus is a raster forcemicroscopic measuring apparatus.
 84. An apparatus in accord with claim 1wherein the measuring apparatus includes an optical detection unit. 85.A method in accord with claim 32 wherein measurements of the stated datafrom the measurement apparatus and/or the evaluation unit are back fedinto the measurement procedure, in order to determine control andevaluation parameters for the said procedure.
 86. A method in accordwith claim 32 wherein data acquired by means of the evaluation of theevaluation unit are automatically stored in a data storage memory.
 87. Amethod in accord with claim 86 wherein the predetermined controlparameters and/or existing measurement conditions of the rastermicroscopic measurement are automatically stored in relation to thecorresponding data produced by the evaluation unit in the data storagememory unit.
 88. A method in accord with claim 32 wherein forces actingupon the probe are evaluated by the evaluation unit.
 89. A method inaccord with claim 32 wherein the probe is set into vibration with apredetermined amplitude regulated by the control unit.
 90. A method inaccord with claim 32 wherein an appropriate force to enable the changeof an effective Q-factor for the probe is directed to the said probeunder the control of the control unit.
 91. A method in accord with theclaims 89 wherein effective vibratory motion changes for the probe arecaptured by the evaluation unit.
 92. A method in accord with claim 32wherein the probe is automatically positioned and/or moved in accordwith predetermined probe positioning parameters by the control unit. 93.A method in accord with claim 92 wherein the positioning and or movingof the probe in accord with predetermined probe positioning parametersof the control parameter group are carried out, which include: movementsof the probe for raster scanning of the sample placed on thesample-carrier, movements of the probe in the vertical direction, amaximum duration of time for a contact of the probe with the sampleplaced upon the sample-carrier, a maximum number of contacts of theprobe with the sample placed upon the sample-carrier, a maximum and/or aminimum probe speed for movements of the same relative to the samplewhich is placed on the sample-carrier, a constant probe speed, aconstant rate of change of probe velocity, a maximum and/or a minimumseparating distance between the probe and the sample placed upon thesample-carrier, a predetermined force, to be held constant between theprobe and the sample placed upon the sample-carrier, a maximum and/or aminimum tension of the probe acting upon the sample placed on thesample-carrier, a maximum and/or a minimum compressive force from theprobe acting upon the sample placed on the sample-carrier, a maximumand/or a minimum rate of change of the tensile force exercised by theprobe on the sample placed on the sample-carrier, a maximum and/or aminimum rate of change of the compressive force exercised by the probeon the sample placed on the sample-carrier.
 94. A method in accord withclaim 32 wherein the positions of the probe and/or the movements of theprobe and/or such forces as may be acting upon the probe are detected bya detector unit, and wherein the first detector unit is automaticallyregulated by the control unit, in accord with predetermined detectionparameters.
 95. A method in accord with claim 94, wherein the detectionof positions of the probe and/or movements of the probe and/or forcesacting upon the probe, in accord with predetermined detection parametersof the control parameter group, is carried out in a manner whichincludes: a predetermined rate of detection, and/or a frequency ofoccurrences, with which the detections for positions of the probe and/ormovements of the probe and/or such forces as may be acting upon theprobe is carried out.
 96. A method in accord with claim 94 wherein thedetected positions and/or movements and/or forces are automaticallyevaluated by an evaluation unit.
 97. A method in accord with claim 93wherein the detected positions and/or movements and/or forces areautomatically classified by the evaluation unit.
 98. A method in accordwith claim 32 wherein the sample-carrier, in accord with predeterminedsample-carrier positioning parameters is automatically positioned and/ormoved by means of the sample-carrier positioning unit under regulationof the control unit.
 99. A method in accord with claim 98 wherein thecontrol of the sample-carrier positioning unit in accord withpredetermined sample-carrier positioning unit parameters of the controlparameter group are carried out, which include: movement of thesample-carrier during the rastered scanning of the sample placed uponthe sample-carrier by the probe, a maximum period of time for a contactof the sample placed upon the sample-carrier with the probe, a maximumnumber of contacts of a sample placed upon the sample carrier with theprobe, a maximum and/or a minimum sample-carrier speed for movements ofthe sample-carrier relative to the probe, a maximum and/or a minimumseparating distance between a sample placed on the sample-carrier andthe probe, a predetermined, constant force to be maintained between thesample placed on the sample-carrier and the probe, a maximum and/or aminimum tensile force by the probe exercised on the sample which hasbeen placed on the sample-carrier, a maximum and/or a minimumcompressive farce by the probe, exercised on the sample which has beenplaced on the sample-carrier, a maximum and/or a minimum change oftension rate for a sample placed upon the sample-carrier, wherein thetension has been exercised by the probe, and/or a maximum and/or aminimum rate of change for a compressive force placed upon thesample-carrier, wherein the compression has been exercised by the probe.100. A method in accord with claim 32 wherein, in the case of anavailable fluid which is confined in a sample chamber of the measurementapparatus, into which fluid the sample is immersed, the said fluid isautomatically regulated by the control unit under conditions ofpredetermined fluid parameters.
 101. A method in accord with claim 100wherein the control of the fluid parameters is in accord with thecontrol parameter group, which encompass the following conditions: apredetermined temperature, a predetermined temperature curve, apredetermined pH value, a predetermined electrolyte content, apredetermined electrolyte content curve, a predetermined flow, apredetermined change of flow, a predetermined fluid level, and/or apredetermined quantity of biological and/or chemical marker features.102. A method in accord with claim 100 wherein, by means of a first feedunit of the sample chamber, fluid, under the regulation of the controlunit is automatically so introduced, that in the case of fluid in thesaid sample chamber predetermined boundary conditions are establishedand maintained.
 103. A method in accord with claim 102 wherein by meansof a second detector unit a liquid level is maintained in the samplechamber and wherein in response to a liquid level detected by the seconddetector unit, the first feed unit is automatically regulated by thecontrol unit.
 104. A method in accord with claim 32 wherein atemperature enclosure, which encases the probe and the sample-carrier isregulated by the control unit under predetermined temperatureparameters.
 105. A method in accord with claim 104 wherein thetemperature enclosure is so controlled, that a predetermined temperatureis held constant or at least a predetermined temperature curve isfollowed.
 106. A method in accord with claim 32 wherein, as a rastermicroscopic measurement, a raster-force microscope measuring operationis carried out.